邻苯二甲醛柱前衍生化高效液相色谱法测定组织中肌肽含量  

Determination of carnosine by reversed phased high performance liquid chromatography with precolumn O-phthalaldehyde derivation

作　　者：杨菁[1] 吴国强[1] 白剑[1] 樊淼[1] 刘春娜[1] 王洪新[1] 

机构地区：[1]辽宁医学院药物研究所,辽宁锦州121001

出　　处：《中国生化药物杂志》2009年第4期258-260,共3页Chinese Journal of Biochemical Pharmaceutics

基　　金：辽宁省自然科学基金(20072202);辽宁省教育厅资助课题(20060528)

摘　　要：目的建立测定小鼠各组织肌肽含量的高效液相色谱法。方法采用c18色谱柱，以甲醇和0．05mol／L磷酸盐缓冲液（pH3．5）为流动相，用邻苯二甲醛为柱前衍生剂，在柱温38℃下采用二元梯度洗脱，对小鼠各组织中的肌肽含量进行了测定。结果肌肽在0．63—10．00μg／mL时峰面积与进样质量浓度有良好的线性关系，其相关系数为0．9996。回收率在98．2％～99．7％之间。检测限为0．05pg／mL（S／N=10）。结论该法灵敏可靠、专属性强、重现性好。小鼠心脏、肝脏、肾脏、大脑皮层和海马中肌肽含量分别为123．67，22．86，158．86，76．57，81．86mg／g。Purpose To establish HPLC method for the determination of carnosine in tissue. Methods The sample was derived with O-phthalaldehyde （OPA）, and then the derivative sample was separated on Kromail C18 column at 38 ℃. A linear gradient elution of A （methanol） and B （0.05 mol/L phosphate buffer,pH 3.5） was used. The flow rate was 1 mL/min, and detected by fluorescence detector with excitation wavelength （λex） at 338 nm and emission wavelength （λem） at 425 nm. Results The calibration curve was liner within the ranges of 0.63-10.00 μg/mL. The correlation coefficient was 0. 999 6. The recoveries were between 98.2%- 99.7%. Detection limit of carnosine was 0.05 μg/mL（S/N = 10）. Conclusion The result shows that this method is simple, rapid, specific, accurate and reproducible. The contents of Carnosine in rat＇s heart, liver, kidney, cortex and hippocampus were 123.67,22.86,158.86,76.57 and 81.86 mg/g （ wet weight）.

关 键 词：肌肽 高效液相色谱法 荧光检测 邻苯二甲醛 含量测定 

分 类 号：Q593[生物学—生物化学]